微小RNA663b对口腔鳞状细胞癌细胞迁移、侵袭和上皮间充质转化的影响

doi:10.7518/hxkq.2022.04.003

华西口腔医学杂志 ›› 2022, Vol. 40 ›› Issue (4): 386-393.doi: 10.7518/hxkq.2022.04.003

微小RNA663b对口腔鳞状细胞癌细胞迁移、侵袭和上皮间充质转化的影响

丛碧桥¹^,²(), 刘晓萍¹, 陈嘉雯¹, 李洪利³, 范欣¹^,³()

^1.潍坊医学院附属医院口腔科，潍坊 261000
^2.潍坊医学院口腔医学院，潍坊 261053
^3.潍坊医学院医学研究实验中心，潍坊 261053

收稿日期:2022-03-23 修回日期:2022-06-10 出版日期:2022-07-25 发布日期:2022-07-27
通讯作者: 范欣 E-mail:cbq20200202@163.com;fanxinsunny@aliyun.com
作者简介:丛碧桥，硕士，E-mail：cbq20200202@163.com
基金资助:
国家自然科学基金青年科学基金项目(81702932);潍坊医学院附属医院种子基金资助(2021wffyzzjj06)

Effect of microRNA-663b on migration, invasion and epithelial‑mesenchymal transition of oral squamous cell carcinoma cells

Cong Biqiao¹^,²(), Liu Xiaoping¹, Chen Jiawen¹, Li Hongli³, Fan Xin¹^,³()

^1.Dept. of Stomatology, Affiliated Hospital of Weifang Medical University, Weifang 261000, China
^2.School of Stomatology, Weifang Medical University, Weifang 261053, China
^3.Medicine Research Center, Weifang Medical University, Weifang 261053, China

Received:2022-03-23 Revised:2022-06-10 Online:2022-07-25 Published:2022-07-27
Contact: Fan Xin E-mail:cbq20200202@163.com;fanxinsunny@aliyun.com
Supported by:
The National Natural Science Foundation of China(81702932);Fundamental Research Program Funding of Affiliated Hospital of Weifang Medical College(2021wffyzzjj06);Correspondence: Fan Xin, E-mail: fanxinsunny@ aliyun.com

摘要/Abstract

摘要：

目的　探讨微小RNA663b（miR-663b）对口腔鳞状细胞癌（OSCC）细胞迁移、侵袭和上皮间质转化（EMT）的影响。方法　通过基因表达数据库（GEO）使用R Studio进行OSCC组织与癌旁正常组织的差异表达分析。实时荧光定量聚合酶链反应（qRT-PCR）检测miR-663b在组织和细胞中的表达。检测miR-663b敲除质粒的HN30细胞转染效率，Transwell法检测迁移、侵袭能力。生物信息学方法预测miR-663b的靶向mRNA，双荧光素酶实验验证结合情况。Western blot检测EMT相关标志物的表达。结果　miR-663b在OSCC组织中表达上调，在HN30、CAL27、SCC-9细胞中表达较HOEC细胞高（P<0.05）。敲除miR-663b可以抑制HN30细胞的迁移和侵袭（P<0.05），抑制EMT的发生。生信预测SH3BP2与miR-663b靶向结合，SH3BP2低表达的患者预后较差（P<0.05）。双荧光素酶实验证明miR-663b可以与SHBP2靶向结合（P<0.05）。敲除miR-663b的OSCC细胞中SH3BP2的表达增加，EMT的发生受到抑制。结论　敲除miR-663b可靶向调节SH3BP2抑制OSCC细胞的迁移、侵袭和EMT。

关键词: miR-663b, SH3BP2, 口腔鳞状细胞癌, 迁移, 侵袭, 上皮间质转化

Abstract:

Objective To explore the effect of microRNA-663b (miR-6636) on migration, invasion and epithelial-mesenchymal transition (EMT) of oral squamous cell carcinoma cells (OSCC). Methods Use R Studio of gene expression omnibus (GEO) database to analyze expressions of miR-663b in the OSCC and adjacent normal tissues. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-663b in tissues and cells. The transfection efficiency of HN30 cells with miR-663b knockout plasmid was detected. Transwell assay was used to detect the effect of the migration and invasion ability. Bioinformatics method was used to predict the targeted mRNA that may bind to miR-663b and double luciferase assay was used to verify the binding. Western blot assay was used to detect the expression of EMT-related markers. Results The expression of miR-663b was up-regulated in OSCC tissues and higher in HN30, CAL27 and SCC-9 cells than in HOEC cells (P<0.05). Knockout of miR-663b could inhibit migration and invasion of HN30 cells (P<0.05) and inhibit the occurrence of EMT. Bioinformatics prediction software predicts that SH3BP2 was the target gene of miR-663b, and patients with low SH3BP2 expression had a poor prognosis (P<0.05). MiR-663b could bind to SHBP2 (P<0.05). The expression of SH3BP2 was increased and the occurrence of EMT was inhibited in HN30 cells with miR-663b knocked out. Conclusion Knockout of miR-663b can inhibit the migration, invasion and EMT of OSCC by targeting SH3BP2.

Key words: miR-663b, SH3BP2, oral squamous cell carcinoma, migration, invasion, epithelial-mesenchymal transition

中图分类号:

R739.8

丛碧桥, 刘晓萍, 陈嘉雯, 李洪利, 范欣. 微小RNA663b对口腔鳞状细胞癌细胞迁移、侵袭和上皮间充质转化的影响[J]. 华西口腔医学杂志, 2022, 40(4): 386-393.

Cong Biqiao, Liu Xiaoping, Chen Jiawen, Li Hongli, Fan Xin. Effect of microRNA-663b on migration, invasion and epithelial‑mesenchymal transition of oral squamous cell carcinoma cells[J]. West China Journal of Stomatology, 2022, 40(4): 386-393.

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微小RNA663b对口腔鳞状细胞癌细胞迁移、侵袭和上皮间充质转化的影响

Effect of microRNA-663b on migration, invasion and epithelial‑mesenchymal transition of oral squamous cell carcinoma cells

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